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<h4 class="current" id="try">Biofilm</h4>

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<h1>Results</h1>
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<h1>Microscopy Images</strong></h1>
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<p style="text-align: center;"><img
	src="http://igem.org/wiki/images/2/25/Zju-biofilm06.jpg" width=660; /></p>
<blockquote>
<p id="ref"><strong>Figure 1</strong><br />
The figure shows the modified DH5alpha’s biofilm formed on rubber tube.
The pink color was due to the RFP expressed by the bacteria. The picture
was obtained with Leica DME microscope, under 40X objective.</p>
</blockquote>
<p style="text-align: center;"><img
	src="Zju-glass1.0.gif" width=660; /></p>
<blockquote>
<p id="ref"><strong>Figure 2</strong><br />
3D CLSM picture of the biofilm formed by E. Coli DH5alpha. The picture
was obtained with Zeiss LSM 710NLO confocal laser scanning microscope,
using a 20X objective. The colors of the biofilm were not from the
fluorescence proteins but artificially added in order to give better
resolution.</p>
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	src="http://2011.igem.org/wiki/images/1/14/Zjubiofilm-figure3.png"
	width="660" ;/></p>
<blockquote>
<p id="ref"><strong>Figure 3</strong><br />
CLSM image of the stratified biofilm obtained with the Cell Culture
Plate Method. It was formed by our modified E.Coli expressing device
BBa_K561003 (you can refer to our Rainbofilm module for more
information). The picture was taken by Zeiss LSM510 Meta confocal laser
scanning microscope, using a 63? oil-immersion objective.</p>
</blockquote>
<p style="text-align: center;"><img
	src="http://2011.igem.org/wiki/images/8/8d/Zju-biofilm07.png"
	width="660" /></p>
<p><strong>Price comparison</strong><br />
Biofilm reactor from biosurface company CDC Biofilm Reactor: $770 Model
DFR 110 Drip Flow Reactor: $989.00 BST Model DK 20-1 Disk Reactor System
with stir plate and support stand: $1,040.00.</p>
<p>&nbsp;</p>
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<h1>Summary</h1>
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<p>The modified DH5alpha biofilm formed on several kinds of
substratum in our experiment, such as rubber tube, cell culture plate,
glass slide and filter membrane. The biofilm, consisting of aggregated
bacteria, was closely anchored on the substratum. Confocal laser
scanning microscopy is a frequently-used and effective tool for biofilm
observation. The profile of biofilm could be clearly observed when RFP
was expressed by the bacteria and the uneven surface structure is shown
in the 3D CLSM picture.</p>
<p>Three methods used to observe the formation of the biofilm were
quite efficient: bubbling method, rubber tube method and cell culture
plate method. It reached the largest thickness in rubber tube assay. In
bubbling assay, the stability of biofilm increased while it was much
thinner. Both the two make use sheer force to accelerate biofilm
formation. In cell culture plate assay, different materials of
substratum were used, including glass slip, PETE and PC. On glass slip,
biofilm maintained the stability and obtained medium thickness that fit
well for further application. What’s more, the unit cost little and the
material helped to achieve a clear image. So we took glass slip laying
in cell culture plate assay as the major method for biofilm formation.</p>
<p>Biofilm formed in two or three days in LB medium. Meanwhile, the
oxygen concentration gradient formed due to the variation of the depth.
According to the modeling results, the gradient became shaper and shaper
during the formation of biofilm, which underlay the construction of a
favorable device for biofilm stratification.</p>

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<p>&nbsp;</p>

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<h1>References</h1>
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<p>1.Bandara HM, Yau JY, Watt RM, Jin LJ, Samaranayake LP.
Escherichia coli and its lipopolysaccharide modulate in vitro Candida
biofilm formation. J Med Microbiol. 2009 Dec; 58(Pt 12): 1623-31. <br />
2. Klausen M, Heydorn A, Ragas P, Lambertsen L, Aaes-Jorgensen A, Molin
S, Tolker-Nielsen T. Biofilm formation by Pseudomonas aeruginosa wild
type, flagella and type IV pili mutants. Mol Microbiol. 2003 Jun; 48(6):
1511-24. <br />
3.Almeida C, Azevedo NF, Santos S, Keevil CW, Vieira MJ. Discriminating
multi-species populations in biofilms with peptide nucleic acid
fluorescence in situ hybridization (PNA FISH). PLoS One. 2011 Mar 29;
6(3): e14786. <br />
4.Teal TK, Lies DP, Wold BJ, Newman DK. Spatiometabolic stratification
of Shewanella oneidensis biofilms. Appl Environ Microbiol. 2006 Nov;
72(11): 7324-30. <br />
5.Rani SA, Pitts B, Beyenal H, Veluchamy RA, Lewandowski Z, Davison WM,
Buckingham-Meyer K, Stewart PS. Spatial patterns of DNA replication,
protein synthesis, and oxygen concentration within bacterial biofilms
reveal diverse physiological states. J Bacteriol. 2007 Jun; 189(11):
4223-33. <br />
6.Gonzalez Barrios AF, Zuo R, Hashimoto Y, Yang L, Bentley WE, Wood TK.
Autoinducer 2 controls biofilm formation in Escherichia coli through a
novel motility quorum-sensing regulator (MqsR, B3022). J Bacteriol. 2006
Jan; 188(1): 305-16. <br />
7. Dickschat JS. Quorum sensing and bacterial biofilms. Nat Prod Rep.
2010 Mar; 27(3): 343-69. <br />
8. Lopez D, Vlamakis H, Kolter R. Biofilms. Cold Spring Harb Perspect
Biol. 2010 Jul; 2(7): a000398.</p>
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